title: Resveratrol decreases B-cell lymphoma-2 expression and viability in GH3 pituitary adenoma cells of the rat.
abstract: OBJECTIVE
  Resveratrol is a phytoestrogen with various antiproliferative and proapoptotic effects. This in vitro study aimed to analyze the effect of resveratrol on the viability and expression of modulators of apoptosis in GH3 pituitary adenoma cells of the rat.
  
  
  METHODS
  GH3 cells were incubated with resveratrol concentrations from 20 to 100 μM for 48-72 hours. Cell viability was quantified using a hemocytometer. We assessed the ability of resveratrol to kill GH3 cells by an enzyme-linked immunosorbent assay (ELISA) of nucleosome liberation and by DNA degradation (unidimensional gel electrophoresis). Relative messenger RNA (mRNA) expression of survivin, B-cell lymphoma-2 protein (BCL-2) and BCL-2-associated X protein (BAX) normalized to β2 microglobulin was measured using quantitative real-time polymerase chain reaction (qRT-PCR).
  
  
  RESULTS
  GH3 cell survival significantly decreased with increasing concentrations of resveratrol. In GH3 cells treated with 100 μM resveratrol, ELISA demonstrated a significant rise of nucleosome liberation, which typically occurs during apoptosis. In parallel, gel electrophoresis showed degradation of DNA into random fragments, pointing to a necrotic mode of cell death in most GH3 cells. In GH3 cells treated with 100 μM resveratrol, qRT-PCR detected a significant decrease of BCL-2 mRNA expression and a decrease of survivin mRNA expression, whereas a change of BAX mRNA expression could not be found. The BAX/BCL-2 ratio was significantly increased in GH3 cells after resveratrol treatment.
  
  
  CONCLUSIONS
  Resveratrol reduces GH3 cell viability in a dose-dependent manner by inducing nonapoptotic cell death and apoptosis. Apoptosis in GH3 cells is probably mediated by resveratrol-dependent downregulation of apoptosis inhibitors, namely BCL-2 and possibly survivin. Further investigation of the potential effects of resveratrol on pituitary adenoma cells is warranted.
authors.affiliation: Department of Neurosurgery, Otto von Guericke University, Magdeburg, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Voellger
authors.forename: Elmar
authors.initials: E
authors.lastname: Kirches
authors.forename: Annette
authors.initials: A
authors.lastname: Wilisch-Neumann
authors.forename: Andreas
authors.initials: A
authors.lastname: Weise
authors.forename: Jorge Humberto
authors.initials: JH
authors.lastname: Tapia-Perez
authors.forename: Rosita
authors.initials: R
authors.lastname: Rupa
authors.forename: Christian
authors.initials: C
authors.lastname: Mawrin
authors.forename: Raimund
authors.initials: R
authors.lastname: Firsching
keywords: GH3 cells; phytoestrogens; resveratrol; viability